Cell sorting of Saccharomyces expressing WT GFP

From: Dominik Stoll (dominik.stoll@twinstrand.com)
Date: Fri Sep 21 2001 - 18:36:02 EST


Cell surface display is a powerful tool for protein analysis. As part of a
fusion protein we were able to display WT GFP on the cell surface of Pichia
pastoris. Under the fluorescence microscope the fusion is clearly visible as
green fluorescence halo around the cells. Attempts to detect and sort the
yeast cells on a FACSvantage machine were unsuccessful. We were not able to
see any difference between negative and positive control cells, possibly due
to auto fluorescence.
What kind of settings should be used for the FACS machine and has anybody
successfully sorted yeast cells using the WT GFP from Aquorea victoria?
Thanks for your contributions,

Dominik Stoll, PhD
Senior Scientist
Twinstrand Therapeutics
www.twinstrand.com
8081 Lougheed Mall
Burnaby BC,
Canada  V5A 1W9
tel: (604) 415 7183
fax: (604) 4157185



This archive was generated by hypermail 2b29 : Sun Jan 05 2003 - 19:01:32 EST